The Wolfson Bioimaging Facility supports state-of-the-art multiphoton imaging with its multi-laser Leica MP/CLSM system.
Multiphoton (MP) microscopy allows imaging deeper within tissues and organisms than our other fluorescence techniques. MP uses two longer wavelength/lower energy photons – which penetrates tissue more readily – to excite fluorophores that are excited at lower wavelengths in single photon techniques such as CLSM.
Excitation requires two photons to be absorbed by the sample almost instantaneously and as power densities are only high enough for this to happen in the focal plane, multiphoton excitation provides inherent optical sectioning.
MP can be used to image fluorophores, intrinsic fluorescence and, by second harmonic generation, structural features of tissue components such as collagen and muscle.
Our multiphoton systems is a Leica SP8 tandem scanning system with Spectra Physics Deep See (680-1300nm plus 1040nm) laser for multiphoton excitation and additional lasers for single photon excitation (Argon, 561 and 633nm). An array of external non-descanned detectors (2PMT+2 HyD) for MP imaging, internal detectors for CLSM (1PMT+2HyD) and a transmitted light detector provide flexibility over imaging options.
Recent publications including data acquired with our multiphoton system:
- Lawrence, Kague, Aggleton, Harniman, Roddy, Hammond (2018) The mechanical impact of col11a2 loss on joints; col11a2 mutant zebrafish show changes to joint development and function, which leads to early-onset osteoarthritis. Phil. Trans. Roy. Soc.
- Walas, Nowicki-Osuch, Alibhai, von Linstow Rolof, Coghill, Waterfall, Paton (2018) Inflammatory pathways are central to posterior cerebrovascular artery remodelling prior to the onset of congenital hypertension. J. Cerebral Blood Flow Metab.
- Morris, Cross, Lu, Kadler, Lu, Dallas, Martin (2018) Live imaging of collagen deposition during skin development and repair in a collagen 1-GFP fusion transgenic zebrafish. Dev. Biol.